Advanced: Gene Regulation
Regulation of Gene Expression: Qualitative
Short Description: Check the expression of a gene by isolating total RNA (kit-based), converting it to cDNA, followed by PCR.
Type: Hands-on
Duration: One day
Fees: INR 2596 (INR 2200 + 18% GST)
Category (for payment): MBY Workshop Tier 2
Aimed at: College students (B.Sc. and M.Sc.)
Application Process: Click here for the details
Registration form (This link will be different for each workshop)
Application process (in brief)
Check the above table to figure out whether suitable dates are available or not.
Note down the category of the workshop.
Head to SB Collect to pay the fees. Save the Receipt and the SB Collect Reference number. See here for details.
Fill the above Registration form using the Receipt and the SB Collect Reference number. Choose the workshop date. You will get an email acknowledgement from us. Registration is NOT yet complete.
We verify your payment and send Registration Confirmation Email. Your registration is now complete.
We strongly encourage the applicants to look at the detailed application process before paying the fees and filling the form.
Course Content
1. Basic laboratory practices and RNA handling practices
This will include basic safety precautions, handling of micropipettes and safe disposal of laboratory wastes along with precautions taken while extracting RNA.
To know more about how to handle micropipettes, watch this video or practice using this simulation. For best RNA handling practices before and after extraction, please read the article here.
2. Understanding steps in RNA isolation from bacteria
Total RNA is isolated using silica column based technique. To know more about general method of RNA isolation, watch this video.
3. Total RNA estimation by spectrophotometry
After completing RNA extraction , the amount of total RNA has to be quantified. The simplest way to do this is through the technique of spectrophotometry which detects the concentration of the final reaction product. To know more about spectrophotometry, you can read this article or watch this video for how a spectrophotometer works.
4. Agarose Gel Electrophoresis (AGE)
The quality of extracted RNA needs to be checked. For this, one needs to perform the technique of gel electrophoresis. To know more about gel electrophoresis, you can read this article or watch this video.
5. Synthesis of cDNA by reverse transcription
Total RNA is used to synthesize complementary DNA (cDNA) using MMLV reverse transcriptase enzyme. The first strand cDNA is then used as a template for gene specific PCR. To know more watch this video.
6. Polymerase chain reaction (PCR)
Once the entire RNA has been converted cDNA, often a specific gene of interest needs to be isolated from it. The simplest way to do this is through the technique of Polymerase Chain Reaction (PCR) which amplifies the number of copies of the gene of interest. To know more about PCR, you can either read this article or watch this video.